LOCUS       sequence-436902-        7417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human Sema4D with N terminal flag after signal peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7417)
  AUTHORS   Bessa P, Newman AG, Yan K, Schaub T, Dannenberg R, Lajko D, 
            Eilenberger J, Brunet T, Textoris-Taube K, Kemmler E, Deng P, 
            Banerjee P, Ravindran E, Preissner R, Rosario M, Tarabykin V
  TITLE     Semaphorin heterodimerization in cis regulates membrane targeting 
            and neocortical wiring.
  JOURNAL   Nat Commun. 2024 Aug 16;15(1):7059. doi: 10.1038/s41467-024-51009-1.
  PUBMED    39152101
REFERENCE   2  (bases 1 to 7417)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2024-08-16"; volume: "15(1):7059. doi"; pages: " 
            10.1038/s41467-024-51009-1"
COMMENT     Sequence Label: pCAG-sp-flag-hSema4D
FEATURES             Location/Qualifiers
     source          1..7417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1797..1820
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4439..4458)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4504..4559
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4558..4577)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4920..4936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4920..4936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4933..4955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4944..4960
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4968..4974,4975..4992,4993..4998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5013..5034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5093..5288
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5139..5274
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5294..5428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5331..5350)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5385..5404
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5496..5513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5667..6255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5747..5766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6426..7217,7218..7286))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7049..7068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7287..7391)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagatg aggatgtgca
     1741 cccccattag ggggctgctc atggcccttg cagtgatgtt tgggacagcg atggcagatt
     1801 ataaagatga tgatgataaa tttgcaccca taccccggat cacctgggag cacagagagg
     1861 tgcacctggt gcagtttcat gagccagaca tctacaacta ctcagccttg ctgctgagcg
     1921 aggacaagga caccttgtac ataggtgccc gggaggcggt cttcgctgtg aacgcactca
     1981 acatctccga gaagcagcat gaggtgtatt ggaaggtctc agaagacaaa aaagcaaaat
     2041 gtgcagaaaa ggggaaatca aaacagacag agtgcctcaa ctacatccgg gtgctgcagc
     2101 cactcagcgc cacttccctt tacgtgtgtg ggaccaacgc attccagccg gcctgtgacc
     2161 acctgaactt aacatccttt aagtttctgg ggaaaaatga agatggcaaa ggaagatgtc
     2221 cctttgaccc agcacacagc tacacatccg tcatggttga tggagaactt tattcgggga
     2281 cgtcgtataa ttttttggga agtgaaccca tcatctcccg aaattcttcc cacagtcctc
     2341 tgaggacaga atatgcaatc ccttggctga acgagcctag tttcgtgttt gctgacgtga
     2401 tccgaaaaag cccagacagc cccgacggcg aggatgacag ggtctacttc ttcttcacgg
     2461 aggtgtctgt ggagtatgag tttgtgttca gggtgctgat cccacggata gcaagagtgt
     2521 gcaaggggga ccagggcggc ctgaggacct tgcagaagaa atggacctcc ttcctgaaag
     2581 cccgactcat ctgctcccgg ccagacagcg gcttggtctt caatgtgctg cgggatgtct
     2641 tcgtgctcag gtccccgggc ctgaaggtgc ctgtgttcta tgcactcttc accccacagc
     2701 tgaacaacgt ggggctgtcg gcagtgtgcg cctacaacct gtccacagcc gaggaggtct
     2761 tctcccacgg gaagtacatg cagagcacca cagtggagca gtcccacacc aagtgggtgc
     2821 gctataatgg cccggtaccc aagccgcggc ctggagcgtg catcgacagc gaggcacggg
     2881 ccgccaacta caccagctcc ttgaatttgc cagacaagac gctgcagttc gttaaagacc
     2941 accctttgat ggatgactcg gtaaccccaa tagacaacag gcccaggtta atcaagaaag
     3001 atgtgaacta cacccagatc gtggtggacc ggacccaggc cctggatggg actgtctatg
     3061 atgtcatgtt tgtcagcaca gaccggggag ctctgcacaa agccatcagc ctcgagcacg
     3121 ctgttcacat catcgaggag acccagctct tccaggactt tgagccagtc cagaccctgc
     3181 tgctgtcttc aaagaagggc aacaggtttg tctatgctgg ctctaactcg ggcgtggtcc
     3241 aggccccgct ggccttctgt gggaagcacg gcacctgcga ggactgtgtg ctggcgcggg
     3301 acccctactg cgcctggagc ccgcccacag cgacctgcgt ggctctgcac cagaccgaga
     3361 gccccagcag gggtttgatt caggagatga gcggcgatgc ttctgtgtgc ccggataaaa
     3421 gtaaaggaag ttaccggcag cattttttca agcacggtgg cacagcggaa ctgaaatgct
     3481 cccaaaaatc caacctggcc cgggtctttt ggaagttcca gaatggcgtg ttgaaggccg
     3541 agagccccaa gtacggtctt atgggcagaa aaaacttgct catcttcaac ttgtcagaag
     3601 gagacagtgg ggtgtaccag tgcctgtcag aggagagggt taagaacaaa acggtcttcc
     3661 aagtggtcgc caagcacgtc ctggaagtga aggtggttcc aaagcccgta gtggccccca
     3721 ccttgtcagt tgttcagaca gaaggtagta ggattgccac caaagtgttg gtggcatcca
     3781 cccaagggtc ttctccccca accccagccg tgcaggccac ctcctccggg gccatcaccc
     3841 ttcctcccaa gcctgcgccc accggcacat cctgcgaacc aaagatcgtc atcaacacgg
     3901 tcccccagct ccactcggag aaaaccatgt atcttaagtc cagcgacaac cgcctcctca
     3961 tgtccctctt cctcttcttc tttgttctct tcctctgcct ctttttctac aactgctata
     4021 agggatacct gcccagacag tgcttgaaat tccgctcggc cctactaatt gggaagaaga
     4081 agcccaagtc agatttctgt gaccgtgagc agagcctgaa ggagacgtta gtagagccag
     4141 ggagcttctc ccagcagaat ggggagcacc ccaagccagc cctggacacc ggctatgaga
     4201 ccgagcaaga caccatcacc agcaaagtcc ccacggatag ggaggactca cagaggatcg
     4261 acgacctttc tgccagggac aagccctttg acgtcaagtg tgagctgaag ttcgctgact
     4321 cagacgcaga tggagactga cctcgaggat atcgcggccg cactcctcag gtgcaggctg
     4381 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     4441 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     4501 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     4561 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     4621 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     4681 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     4741 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     4801 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     4861 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     4921 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4981 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5041 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     5101 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5161 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5221 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5281 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5341 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5401 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     5461 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5521 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6301 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6541 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6721 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7021 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7381 ggggttccgc gcacatttcc ccgaaaagtg ccacctg
//
